Department of Cellular and Molecular Medicine, Faculty of Health and Medical Sciences, Copenhagen Center for Glycomics, University of Copenhagen, Copenhagen, Denmark.
Methods Mol Biol. 2022;2442:205-214. doi: 10.1007/978-1-0716-2055-7_12.
The family of galectins has critical functions in a wide range of biological processes, primarily based on their broad interactions with proteins carrying β-galactoside-containing glycans. To understand the diversity of functions governed by galectins, it is essential to define the binding specificity of the carbohydrate recognition domain (CRDs) of the individual galectins. The binding specificity of galectins has primarily been examined with glycoarrays, but now the ability to probe and dissect binding to defined glycans in the context of a cellular membrane is facilitated by the generations of glycoengineered cell libraries with defined glyco-phenotypes. The following section will show how galectin specificities can be probed in the natural context of cellular surfaces using glycoengineered cell libraries, and how binding to glycoproteins can be measured in solution with fluorescence anisotropy.
半乳糖凝集素家族在广泛的生物学过程中具有关键功能,主要基于其与携带β-半乳糖苷聚糖的蛋白质的广泛相互作用。为了了解半乳糖凝集素所调控的功能多样性,定义各个半乳糖凝集素的碳水化合物识别结构域 (CRD) 的结合特异性至关重要。半乳糖凝集素的结合特异性主要通过糖组阵列进行研究,但现在通过生成具有定义糖表型的糖工程化细胞文库,可以在细胞膜的背景下探测和剖析与特定糖的结合,从而使这一能力成为可能。以下部分将展示如何使用糖工程化细胞文库在细胞表面的自然环境中探测半乳糖凝集素的特异性,以及如何通过荧光各向异性测量溶液中糖蛋白的结合。